Protein precipitation - acetone?
(by Nikola.Wenta from nottingham.ac.uk)
Thu Mar 11 12:27:40 EST 2010
I don't have any clue about acetone precipitation of proteins, but generally, precipitation with saturated Ammonium sulfate solution provides a good means to concentrate proteins and to get them into a save state for short-term storage. Unfortunatelly, you would already need the protein solution to be at > 1 mg/ml in order to get it to precipitate. Thus, in your case this method doesn't seem suitable. You could also try to concentrate the protein with Centricons, but you would rather loose protein to the membrane than concentrate your solution as it is already too diluted. Why not loading maximum volume into biggest possible pockets on a gel with maximum thick spacers? Additionally you could use a acrylamide percentage that "compresses" your protein band, giving you a better signals in WB.
Von: methods-bounces from oat.bio.indiana.edu im Auftrag von methods-request from oat.bio.indiana.edu
Gesendet: Do 11.03.2010 17:03
An: methods from magpie.bio.indiana.edu
Betreff: Methods Digest, Vol 58, Issue 7
Date: Thu, 11 Mar 2010 16:56:05 +0100
From: "Iraz Toprak Aydin" <iraz.aydin from epfl.ch>
Subject: Protein precipitation - acetone?
To: <methods from magpie.bio.indiana.edu>
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I have to do a western blot, but my protein concentration is very low, and I
have to run a mini gel. So I was thinking of precipitation the proteins. I
have never done this before. Does acetone have a bad effect on the blotting?
Are there any points that I should be careful about?
Thanks in advance...
Iraz Toprak Aydin
EPFL SV ISREC, Station 19
Batiment SV, SV 2540
Tel: +41 21 693 07 36
e-mail: <mailto:iraz.aydin from epfl.ch> iraz.aydin from epfl.ch
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